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Culture medium for culturing fat stem cells and preparation method thereof

A technology of adipose stem cells and culture medium, applied in biochemical equipment and methods, culture process, tissue culture, etc., can solve the problems of complicated culture methods, discomfort of stem cells, easy aging, etc., shorten the culture time, maintain cell viability, improve safety effect

Inactive Publication Date: 2018-03-23
沈国青
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the prior art, after continuous culture in vitro, adipose stem cells are prone to aging, karyotype changes, surface antigen changes, and multidirectional differentiation ability is limited to osteoblasts or osteoblasts and adipocytes. The method is complicated, the number of extracted stem cells is small, the purity is not high, and the subculture proliferation is slow. What's more, the use of feeder cells and animal serum is easy to cause stem cell contamination, especially the potential animal-derived endotoxin or virus in animal serum will be harmful to the human body. Health poses a great risk. The stem cells cultured in this way are not suitable for direct clinical application, which limits their research and application in adipose tissue engineering and other tissue engineering.

Method used

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Examples

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Comparison scheme
Effect test

Embodiment 1

[0026] This embodiment provides a medium for culturing adipose stem cells, including basal medium and additives, in parts by mass, and the additives include the following substances:

[0027] 7 parts of soybean peptone, 3 parts of sodium chloride, 2 parts of glucose, 2.6 parts of lactose, 1.8 parts of phosphorus source, 0.08 parts of pig bile salt, 1.6 parts of mannitol, 1.8 parts of sodium pyruvate, 2.3 parts of glycine, 1.9 parts of vitamins, β - 1.2 parts of lactam antibiotics, 0.43 parts of iron salt, 0.27 parts of wolfberry polysaccharide, 1000 parts of distilled water;

[0028] This embodiment also provides the preparation method of the medium for culturing adipose stem cells, comprising the following steps:

[0029] (1) Dissolve the dry powder of DMEM:F12 in water to obtain the basal medium;

[0030] (2) Add soybean peptone, sodium chloride, glucose, lactose, phosphorus source, pig gallbladder, mannitol, sodium pyruvate, glycine, vitamins, β-lactam antibiotics, iron sa...

Embodiment 2

[0033] This embodiment provides a medium for culturing adipose stem cells, including basal medium and additives, in parts by mass, and the additives include the following substances:

[0034]5 parts of blood peptone, 2 parts of sodium chloride, 1 part of glucose, 0.8 parts of lactose, 1.2 parts of phosphorus source, 0.05 parts of pig bile salt, 0.3 parts of mannitol, 0.8 parts of sodium pyruvate, 0.5 parts of L-glutamine, vitamin 0.5 parts, 0.5 parts of aminoglycoside antibiotics, 0.12 parts of copper salt, 0.23 parts of astragalus polysaccharide, 1000 parts of distilled water;

[0035] This embodiment also provides the preparation method of the medium for culturing adipose stem cells, comprising the following steps:

[0036] (1) Dissolve the dry powder of DMEM:F12 in water to obtain the basal medium;

[0037] (2) Add blood peptone, sodium chloride, glucose, lactose, phosphorus source, pig gallbladder, mannitol, sodium pyruvate, L-glutamine, vitamins, aminoglycoside antibioti...

Embodiment 3

[0040] This embodiment provides a medium for culturing adipose stem cells, including basal medium and additives, in parts by mass, and the additives include the following substances:

[0041] 12 parts of tryptone, 6 parts of sodium chloride, 3 parts of glucose, 5.6 parts of lactose, 2.8 parts of phosphorus source, 0.1 part of pig bile salt, 2.1 parts of mannitol, 2.0 parts of sodium pyruvate, 2.1 parts of L-arginine, vitamin 2.3 parts, 1.2 parts of polypeptide antibiotics, 0.36 parts of metal salts, 0.44 parts of Morinda officinalis polysaccharide, 1000 parts of distilled water;

[0042] This embodiment also provides the preparation method of the medium for culturing adipose stem cells, comprising the following steps:

[0043] (1) Dissolve the dry powder of DMEM:F12 in water to obtain the basal medium;

[0044] (2) Add tryptone, sodium chloride, glucose, lactose, phosphorus source, pig gallbladder, mannitol, sodium pyruvate, L-arginine, vitamins, polypeptide antibiotics, meta...

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Abstract

The invention relates to the technical field of microculture, in particular to a culture medium for culturing fat stem cells and a preparation method thereof. The culture medium comprises a basic culture medium and additives, and the additives include peptone, sodium chloride, glucose, lactose, phosphorus source, pig gall salt, mannitol, sodium pyruvate, amino acid, vitamin, antibiotic, metal salt, plant polysaccharide and distilled water. The culture medium contains the plant polysaccharide, thereby being capable of inhibiting growth of other cells except for fat stem cells and promoting proliferation of the fat stem cells and conducive to increasing concentration of the fat stem cells in the culture medium and shortening culture time; through synergism among ingredients, sufficient nutrition and good environment needed for cell growth and proliferation can be provided, cell growth can be promoted, albumin synthesis can be promoted, and cell activity can be maintained for a long time.

Description

technical field [0001] The invention relates to the technical field of microorganism culture, in particular to a medium for culturing fat stem cells and a preparation method thereof. Background technique [0002] Adipose-derived stem cells (ADSCs) are a kind of stem cells with multi-directional differentiation potential isolated from adipose tissue in recent years. They have the characteristics of general stem cells such as rapid expansion and not easy to age. Human adipose stem cells, namely human adipose-derived mesenchymal stem cells, are a kind of adult stem cells widely used in the field of tissue engineering and regenerative medicine, which have the same multi-directional differentiation potential as bone marrow mesenchymal stem cells. ADSCs showed fibroblast-like growth, abundant cytoplasm and nucleoli, arranged in parallel or swirl-like arrangement. Cell cycle analysis showed that 69% of cells were in G0 / G1 phase, 24% in S phase, and 8% in G2 / M phase. The cells wer...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2500/12C12N2500/30C12N2500/32C12N2500/34C12N2500/35C12N2500/38
Inventor 沈国青
Owner 沈国青
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